Feasibility of ultra-high performance liquid and gas chromatography coupled to mass spectrometry for accurate determination of primary and secondary phthalate metabolites in urine samples

•GC-EI-ITD(SIM) has been validated to determine 9 MPAEs in urine samples.•UHPLC-ESI(−)QqQ MS2 has been validated to determine 9 MPAEs in urine samples.•9 MPAEs were determined in normal Spanish human urine samples.•Differences in MPAE urine levels were found between Spanish men and women.

Phthalates (PAEs) are ubiquitous toxic chemical compounds. During the last few years, some phthalate metabolites (MPAEs) have been proposed as appropriate biomarkers in human urine samples to determine PAE human intake and exposure. So, it is necessary to have fast, easy, robust and validated analytical methods to determine selected MPAEs in urine human samples. Two different instrumental methods based on gas (GC) and ultra-high performance liquid (UHPLC) chromatography coupled to mass spectrometry (MS) have been optimized, characterized and validated for the simultaneous determination of nine primary and secondary phthalate metabolites in urine samples. Both instrumental methods have similar sensitivity (detection limits ranged from 0.03 to 8.89 pg μL−1 and from 0.06 to 0.49 pg μL−1 in GC–MS and UHPLC–MS2, respectively), precision (repeatability, expressed as relative standard deviation, which was lower than 8.4% in both systems, except for 5OH-MEHP in the case of GC–MS) and accuracy. But some advantages of the UHPLC–MS2 method, such as more selectivity and lower time in the chromatographic runs (6.8 min vs. 28.5 min), have caused the UHPLC–MS2 method to be chosen to analyze the twenty one human urine samples from the general Spanish population. Regarding these samples, MEP showed the highest median concentration (68.6 μg L−1), followed by MiBP (23.3 μg L−1), 5cx-MEPP (22.5 μg L−1) and MBP (19.3 μg L−1). MMP (6.99 μg L−1), 5oxo-MEHP (6.15 μg L−1), 5OH-MEHP (5.30 μg L−1) and MEHP (4.40 μg L−1) showed intermediate levels. Finally, the lowest levels were found for MBzP (2.55 μg L−1). These data are within the same order of magnitude as those found in other similar populations.

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